Binding of Acanthamoeba to [corrected] mannose-glycoproteins of corneal epithelium: effect of injury.

Abstract:

PURPOSE:Acanthamoeba keratitis is a sight-threatening corneal infection. It is known that: (i) more amoebae bind to the surface of injured corneas than to the normal corneal surface and (ii) mannose-containing glycoproteins (GPs) possess binding sites for Acanthamoeba. The present study was undertaken to determine whether subtle corneal surface injury exposes mannose-GPs and whether more amoebae bind to the mannose-GPs of injured corneas than to those of normal corneas. METHODS:Corneal cup assays were developed to determine whether corneal surface injury exposes binding sites for a mannose/glucose-specific lectin, succinylated-concanavalin A (s-ConA). To determine whether injury exposes mannose-GPs, corneal surface proteins were biotinylated, biotin-labeled mannose-GPs were allowed to bind to s-ConA-agarose beads and were analyzed by SDS-polyacrylamide gel electrophoresis (PAGE). Amoeba binding to mannose-GPs of corneal epithelia was analyzed by PAGE-blot overlay assays. RESULTS:S-ConA binding site density was 2.4 times greater on the injured corneal surface than on the surface of normal corneas. Based on the analysis of the s-ConA-bound, biotin-labeled corneal surface proteins, approximately 5.2 times greater amounts of mannose-GPs were present on the surface of injured corneas than on the normal corneal surface. PAGE-blot overlay assays of s-ConA bound GPs of unlabeled corneal epithelia revealed that, on a per mg total cell protein basis, injured corneal epithelium contained 1.8 times greater amounts of Acanthamoeba-reactive mannose-GPs than normal corneal epithelium. CONCLUSIONS:Subtle corneal injury exposes mannose-GPs on the surface of injured corneas. The newly exposed GPs may serve to provide additional attachment sites for the amoebae. This, in turn, could render the cornea susceptible to the infection.

journal_name

Curr Eye Res

journal_title

Current eye research

authors

Jaison PL,Cao Z,Panjwani N

subject

Has Abstract

pub_date

1998-08-01 00:00:00

pages

770-6

issue

8

eissn

0271-3683

issn

1460-2202

journal_volume

17

pub_type

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